US5650627A - Sensitive assay method for measuring gallium levels in body tissues and fluids - Google Patents
Sensitive assay method for measuring gallium levels in body tissues and fluids Download PDFInfo
- Publication number
- US5650627A US5650627A US08/355,153 US35515394A US5650627A US 5650627 A US5650627 A US 5650627A US 35515394 A US35515394 A US 35515394A US 5650627 A US5650627 A US 5650627A
- Authority
- US
- United States
- Prior art keywords
- test sample
- assay method
- absorption
- gallium
- zeeman
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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- GYHNNYVSQQEPJS-UHFFFAOYSA-N Gallium Chemical compound [Ga] GYHNNYVSQQEPJS-UHFFFAOYSA-N 0.000 title claims abstract description 27
- 229910052733 gallium Inorganic materials 0.000 title claims abstract description 27
- 238000003556 assay Methods 0.000 title claims abstract description 21
- 239000012530 fluid Substances 0.000 title description 5
- 238000012360 testing method Methods 0.000 claims abstract description 31
- 238000010521 absorption reaction Methods 0.000 claims abstract description 25
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229910017604 nitric acid Inorganic materials 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 12
- 238000003705 background correction Methods 0.000 claims abstract description 11
- 210000001124 body fluid Anatomy 0.000 claims abstract description 7
- 239000010839 body fluid Substances 0.000 claims abstract description 6
- 238000000889 atomisation Methods 0.000 claims abstract description 5
- 238000007865 diluting Methods 0.000 claims abstract description 3
- 239000011159 matrix material Substances 0.000 claims description 3
- 210000002700 urine Anatomy 0.000 description 6
- 238000003321 atomic absorption spectrophotometry Methods 0.000 description 5
- 238000001479 atomic absorption spectroscopy Methods 0.000 description 5
- CHPZKNULDCNCBW-UHFFFAOYSA-N gallium nitrate Chemical compound [Ga+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O CHPZKNULDCNCBW-UHFFFAOYSA-N 0.000 description 4
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- WTDHULULXKLSOZ-UHFFFAOYSA-N Hydroxylamine hydrochloride Chemical compound Cl.ON WTDHULULXKLSOZ-UHFFFAOYSA-N 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 239000013060 biological fluid Substances 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 229960004316 cisplatin Drugs 0.000 description 2
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 2
- 229940044658 gallium nitrate Drugs 0.000 description 2
- 230000003211 malignant effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000000559 atomic spectroscopy Methods 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- 190000008236 carboplatin Chemical compound 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000011254 conventional chemotherapy Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229910002804 graphite Inorganic materials 0.000 description 1
- 239000010439 graphite Substances 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000007479 molecular analysis Methods 0.000 description 1
- 238000009520 phase I clinical trial Methods 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 229940043267 rhodamine b Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/3103—Atomic absorption analysis
Definitions
- Kelsen et al. assayed for gallium levels in plasma and urine by flameless atomic absorption spectrophotometry (AAS) using a PERKIN-ELMER 372 atomic absorption spectrophotometer. It was reported that gallium metal (500 mg) was dissolved in 5 ml of concentrated nitric acid and diluted with water to 1 liter, with standards being prepared daily by appropriate dilution.
- the working range was indicated to be 0.1-2.0 micrograms/ml for aqueous standards and 0.1-0.5 micrograms/ml, when 0.5N HNO 3 or plasma were used as diluents. It was reported that gallium levels as low as 0.025 micrograms/ml were measurable in plasma treated and in urine diluted with 0.5N HNO 3 , and that sensitivity was decreased approximately five-fold in aqueous solutions of standards or urine.
- Reed et al. Atomic Spectroscopy, Vol. 9, No. 3, pp. 93-95 (1988), reported that the ability to measure elemental platinum was enhanced by more than a log, using Atomic Absorption Spectroscopy methods with Zeeman background correction. In this respect, a series of tissue culture studies have been performed using cisplatin or carboplatin, using the method reported by Reed et al.
- the present invention provides a highly sensitive assay method for measuring the level of elemental gallium present in body tissues and fluids.
- the assay method employs an atomic absorption spectrometer having Zeeman-effect background correction capabilities.
- gallium levels can be measured in a test sample of body tissue or fluid to a sensitivity of about 25 picograms.
- the inventive assay method more specifically allows one to sensitively measure the quantity of elemental gallium present in a test sample comprising a body tissue or a body fluid by carrying out the steps of:
- test sample's absorption is preferably determined at a wavelength of about 287 nanometers, more preferably about 287.4 nanometers.
- said test sample is diluted in a matrix of less than about 2% (volume/volume) of nitric acid, more preferably less than about 1% (volume/volume) of nitric acid, and most preferably contains about 0.2% (volume/volume) nitric acid.
- the body tissue or fluid test sample to be analyzed is that of a human, and preferably the test sample has a volume of about 50 microliters or less.
- the present inventive assay method has a detection limit of about 25 picograms of elemental gallium, as compared to a calculated 0.5 micrograms (500 picograms) of gallium for the reported method of Kelsen et al., supra., this produces at least two notable advantages:
- the present inventive assay method depends in part to the use of an atomic absorption spectrometer (AAS) having the ability to perform a Zeeman-effect background correction calculation.
- AAS atomic absorption spectrometer
- Such spectrometers are well known in the art and are commercially available.
- such a spectrometer was used by Reed et al. supra., in order to quantify platinum-DNA binding after therapeutic exposure to cisplatin.
- Exemplary of such a spectrometer is the PERKIN-ELMER 3030B, which was used in the Example provided below.
- atomic absorption spectrometer should have the capability to perform a Zeeman-effect background correction calculation, in order to fully enjoy the high degree of sensitivity that is possible with the inventive assay methods. Further, whatever atomic absorption spectrometer is chosen, it should be operated according to the manufacturers instructions, and in accordance with standard atomic absorption spectroscopy techniques utilized by those skilled in the art. In this respect, the following texts on atomic absorption spectroscopy and atomic absorption spectroscopy techniques are mentioned and are incorporated herein by reference, in their entirety:
- the present inventive assay methods are applicable to a variety of body tissues and fluids. For example they are applicable to plasma, urine, blood, etc., of both human and non-human origin. Moreover, because the present inventive methods are highly sensitive to the presence of gallium--they allow one to study the subcellular effects of gallium. As a result, it is possible to use the present assay methods to measure gallium levels in bacteria (e.g., E. coli), yeast, ground tissues and cell cultures, all of which are encompassed by the use of the terms "body tissues” and "body fluids,” as they are used herein and in the accompanying claims.
- bacteria e.g., E. coli
- yeast e.g., yeast, ground tissues and cell cultures
- Sample sizes to be analyzed were 50 microliters, charting temperatures of 130° C., pretreatment at 800° C., atomization at 2400° C., and burn off at 2600° C.
- Standard curves were made using multiple injections of a range of 0.5 to 40 nanograms/ml of elemental gallium in matrices of dH 2 O and 0.2% nitric acid (HNO 3 ).
- the present inventive assay method with using a 0.2% (volume/volume HNO 3 matrix for analysis of gallium permitted the determination of low levels of gallium with very good reproducibility in biological materials.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Hematology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims (6)
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/355,153 US5650627A (en) | 1994-12-08 | 1994-12-08 | Sensitive assay method for measuring gallium levels in body tissues and fluids |
PCT/US1995/015987 WO1996018097A1 (en) | 1994-12-08 | 1995-12-08 | Sensitive assay method for measuring gallium levels in body tissues and fluids |
AU45122/96A AU4512296A (en) | 1994-12-08 | 1995-12-08 | Sensitive assay method for measuring gallium levels in body tissues and fluids |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/355,153 US5650627A (en) | 1994-12-08 | 1994-12-08 | Sensitive assay method for measuring gallium levels in body tissues and fluids |
Publications (1)
Publication Number | Publication Date |
---|---|
US5650627A true US5650627A (en) | 1997-07-22 |
Family
ID=23396429
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US08/355,153 Expired - Lifetime US5650627A (en) | 1994-12-08 | 1994-12-08 | Sensitive assay method for measuring gallium levels in body tissues and fluids |
Country Status (3)
Country | Link |
---|---|
US (1) | US5650627A (en) |
AU (1) | AU4512296A (en) |
WO (1) | WO1996018097A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109883972A (en) * | 2019-04-11 | 2019-06-14 | 哈尔滨工业大学(威海) | Dissociate Methods For The Determination of Iron in a kind of red blood cell |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SU1216726A1 (en) * | 1983-12-29 | 1986-03-07 | Московский Ордена Трудового Красного Знамени Научно-Исследовательский Институт Гигиены Им.Ф.Ф.Эрисмана | Method of intoxication diagnosis by heavy metals |
JPH01302141A (en) * | 1988-05-30 | 1989-12-06 | Shimadzu Corp | Analysis of very small amount of gallium by flameless atomic absorptiometry |
US5106189A (en) * | 1990-03-16 | 1992-04-21 | Hitachi, Ltd. | Zeeman atomic absorption spectrophotometer |
US5225182A (en) * | 1991-10-31 | 1993-07-06 | Sharma Yash P | Vectored drug delivery system using a cephaloplastin linking agent and a methed of using the system |
US5315528A (en) * | 1991-12-03 | 1994-05-24 | The Perkin-Elmer Corporation | Method of, and apparatus for, absorbance correction in atomic absorption spectroscopy |
-
1994
- 1994-12-08 US US08/355,153 patent/US5650627A/en not_active Expired - Lifetime
-
1995
- 1995-12-08 WO PCT/US1995/015987 patent/WO1996018097A1/en active Application Filing
- 1995-12-08 AU AU45122/96A patent/AU4512296A/en not_active Abandoned
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SU1216726A1 (en) * | 1983-12-29 | 1986-03-07 | Московский Ордена Трудового Красного Знамени Научно-Исследовательский Институт Гигиены Им.Ф.Ф.Эрисмана | Method of intoxication diagnosis by heavy metals |
JPH01302141A (en) * | 1988-05-30 | 1989-12-06 | Shimadzu Corp | Analysis of very small amount of gallium by flameless atomic absorptiometry |
US5106189A (en) * | 1990-03-16 | 1992-04-21 | Hitachi, Ltd. | Zeeman atomic absorption spectrophotometer |
US5225182A (en) * | 1991-10-31 | 1993-07-06 | Sharma Yash P | Vectored drug delivery system using a cephaloplastin linking agent and a methed of using the system |
US5315528A (en) * | 1991-12-03 | 1994-05-24 | The Perkin-Elmer Corporation | Method of, and apparatus for, absorbance correction in atomic absorption spectroscopy |
Non-Patent Citations (9)
Title |
---|
Cernik, "A Preliminary Procedure for the Determination of Cadmium in Blood," Atomic Absorption Newsletter, vol. 12, No. 6, 1973, pp. 163-164. |
Cernik, A Preliminary Procedure for the Determination of Cadmium in Blood, Atomic Absorption Newsletter, vol. 12, No. 6, 1973, pp. 163 164. * |
D. P. Kelsen et al., "Pharmacokinetics of Gallium Nitrate in Man," Cancer, vol. 46, 1980, pp. 2009-2013. |
D. P. Kelsen et al., Pharmacokinetics of Gallium Nitrate in Man, Cancer, vol. 46, 1980, pp. 2009 2013. * |
E. Reed et al., "Quantitation of Platinum-DNA Binding After Therapeutic Levels of Drug Exposure--A Novel Use of Graphite Furnace Spectrometry," Atomic Spectroscopy, May-Jun. 1988, vol. 9, No. 3, pp. 93-95. |
E. Reed et al., Quantitation of Platinum DNA Binding After Therapeutic Levels of Drug Exposure A Novel Use of Graphite Furnace Spectrometry, Atomic Spectroscopy, May Jun. 1988, vol. 9, No. 3, pp. 93 95. * |
K. B. Lee, Proceedings of the American Association for Cancer Research, vol. 36, p. 312, 1857 (1995). * |
S. W. Hall et al., "Kinetics of gallium nitrate, a new anticancer agent," Clin Pharmacol. Ther., vol. 25, No. 1, Jan. 1979, pp. 82-87. |
S. W. Hall et al., Kinetics of gallium nitrate, a new anticancer agent, Clin Pharmacol. Ther., vol. 25, No. 1, Jan. 1979, pp. 82 87. * |
Also Published As
Publication number | Publication date |
---|---|
WO1996018097A1 (en) | 1996-06-13 |
AU4512296A (en) | 1996-06-26 |
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Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: GOVERNMENT OF THE UNITED STATES OF AMERICA, THE, A Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:REED, EDDIE;LEE, KANG BO;REEL/FRAME:007336/0909 Effective date: 19950208 |
|
AS | Assignment |
Owner name: DEPARTMENT OF HEALTH AND HUMAN SERVICES, GOVERNMEN Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:REED, EDDIE;LEE, KANG BO;REEL/FRAME:008530/0746;SIGNING DATES FROM 19970423 TO 19970520 |
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Year of fee payment: 4 |
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